摘要
将含有斜带石斑鱼(Epinephelus coioides)神经坏死病毒(orange-spotted grouper nervous necrosis virus,OGNNV)的主衣壳蛋白(main capsid protein,MCP)基因的重组质粒pET32a-MCP转入大肠杆菌BL21后,用异丙基硫代-β-D半乳糖苷(IPTG)诱导表达,用柱层析纯化表达的融合蛋白作为抗原免疫新西兰大白兔,制备抗MCP融合蛋白的血清。用ELISA方法检测抗血清的效价,用Western-blot检测抗血清的特异性。结果显示,获得的抗血清稀释1∶22 000倍时仍呈阳性,并能有效中和OGNNV,实验组的相对存活率达54.50%。这说明,本研究用纯化的MCP融合蛋白制备兔抗OGNNVMCP抗体是成功的,并证实了OGNNV主衣壳蛋白的免疫原性。本研究旨为重组表达主衣壳蛋白基因制备抗OGNNV疫苗提供科学依据,并为进一步研究提供重要的实验材料。
Recombinant expression vector pET32a-MCP which inserted with MCP gene of OGNNV was transformed into BL21 for expression of MCP by inducing of IPTG. Polyclonal antibody to fusion protein was prepared from rabbits immunized with recombinant protein purified through column chromatography.The titre of anti recombinant protein sera was tested by ELISA. Western blot analysis confirmed specificity of the anti-sera. Results showed that the antibodies with the titre of 1：22 000 could effectively neutralize OGNNV, and the relative percentage survival（RPS） of the fish iniected with antibodies reached 54.50 %. The antibodies was successfully prepared with purified recombinant MCP. Conclusion： which confirmed the immunogenicity of MCP from OGNNV and provided the scientific basis for anti-C）ONNV vaccine of recombinant MCP. Also,the polyclonal antibodies can be used in further studies.
出处
《中国水产科学》
CAS
CSCD
北大核心
2006年第5期841-844,共4页
Journal of Fishery Sciences of China
基金
国家高技术研究发展计划(863计划)项目(2003AA603011)
暨南大学校内博士启动基金(51204062)
关键词
斜带石斑鱼
神经坏死病毒
主衣壳蛋白
抗体
orange-spotted grouper nervous necrosis virus
main capsid protein
antibodies
